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Addgene

pET28-MHL
(Plasmid #26096)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 26096 Standard format: Plasmid sent in bacteria as agar stab 1 $85 *

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Backbone

  • Vector backbone
    pET28-MHL
  • Backbone manufacturer
    Structural Genomics Consortium
  • Backbone size (bp) 7325
  • Vector type
    Bacterial Expression
  • Promoter T7-lacO
  • Tag / Fusion Protein
    • Hexahistidine tag with TEV cleavage (N terminal on backbone)

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Low Copy

Cloning Information

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

The pET28-MHL vector (GenBank accession EF456735) was derived from expression plasmid pET28a-LIC (SGC). It is used for T7 promoter driven expression of recombinant proteins with the addition of an 18 amino acid N-terminal fusion tag containing 6X His followed by a TEV cleavage site. The GSS residues after the Met start site were removed to reduce N-terminal gluconoylation via preventing N-terminal Met excision. Two stop codons are included in the vector at the C-terminal cloning site.

Insertion of DNA sequence into the cloning/expression region is preformed using BD-Biosciences Infusion enzyme mediated directional recombination between complementary 15 nucleotide DNA sequences at the ends of the insert (PCR product) and BseRI linearized vector. Insertion of target sequence involves replacement of a SacB gene stuffer sequence, which provides for negative selection of the original plasmid on 5% sucrose.

http://www.sgc.utoronto.ca/SGC-WebPages/toronto-vectors.php

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pET28-MHL was a gift from Cheryl Arrowsmith (Addgene plasmid # 26096 ; http://n2t.net/addgene:26096 ; RRID:Addgene_26096)